In the past few decades, a substantial amount of time and effort has been put into researching the development of female reproductive organs. The impetus behind the research varies from laboratory to laboratory, however, all the research efforts address important common issues relating to the overall health of women. Some of these issues include: cervical cancer, infertility, endometriosis, uterine cancer, ectopic pregnancies, ovarian cysts, and uterine fibroids. Cervical cancer, for example, is a particularly important topic for women's health considering that cervical cancer is the second most common cancer among women worldwide with approximately 450,000 new cases being diagnosed annually and that almost 200,000 deaths are due to cervical cancer. Pisani et al. Int. J. Cancer 55: 891–903 (1993). Although the etiological cause of cervical cancer remains unknown today, there are many reports that infection with human papillomavirus, in particular, HPV-16 and HPV-18, may be the cause for the development of cervical cancer. Although cervical cancer research has accomplished progress in the past, some of the most critical work is impeded by a lack of human tissue models. Likewise, research relating to ovarian cancers, uterine cancer, uterine fibroids, or endometriosis would benefit greatly from having human tissue models of the cervix, uterus, oviduct (fallopian tube), and vagina.
The cervix, uterus, oviduct, and part of the vagina of the female reproductive system are formed early in embryogenesis from Müllerian ducts, also known as paramesonephric ducts. In human embryos, a primordial gonadal ridge develops into a primitive gonad. At about the seventh week of gestation, both sexes have primordial genital ducts and a primitive gonad which develops into a cortex and a medulla. In genetic females, the cortex develops into ovaries and the medulla regresses. In contrast, the medulla develops into testes and the cortex regresses in genetic males. As development of a human embryo progresses, Müllerian ducts in males begin to regress with the secretion of Müllerian inhibiting substance (or MIS). Ganong, William F. Review of Medical Physiology, Chapter 23 “The Gonads: Development and Function of the Reproductive System”, Fifteenth Edition, Appleton and Lange (1991). The Müllerian duct is either of the two paired embryonic tubes extending along the mesonephros roughly parallel to the mesonephric duct and emptying into the cloaca. In females, the upper parts of the Müllerian duct form the oviducts, while the lower parts fuse to form the uterus, cervix, and part of the vagina.
Previous work on Müllerian ducts have focused on anatomical and structural characteristics of Müllerian ducts. For example, one study revealed that the movements of Müllerian ridges and the immunohistochemical staining of Müllerian ducts in avians closely resemble that seen in human. Jacob M, et. al. Cells Tissues Organs 164(2), 63–81, (1999). In another study, human fetuses were examined by ultrasound to study the developing urogenital tracts. Lawrence W. D., et. al. American Journal of Obstetrics and Gynecology 167(1), 185–193, (1992). Other studies have focused on gene expression patterns in the developing fetus. Pellegrini M. et. al. Anat. Embryol. 196(6). 427–433, (1997). While important in their respective scopes, these studies do not provide any teachings for methods of isolating Müllerian duct-derived epithelial cells, nor do they provide any teaching for methods for culturing Müllerian duct-derived epithelial cells such that the cells retain their pluripotent potential. There are very few, if any, reports of Müllerian duct-derived epithelial cells that have been isolated and even fewer reports of Müllerian duct-derived epithelial cells that have pluripotent potential to differentiate into uterine, cervical, oviductal, and vaginal cells. Accordingly, there exists a need for the discovery of a population of Müllerian duct-derived epithelial cells as well as methods of isolating and characterizing Müllerian duct-derived epithelial cells. The invention disclosed herein fulfills these needs and discloses additional methods of use as well.